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泰山医学院毕业设计（论文）题目：蜂胶黄酮抗H2O2诱导PC12细胞

凋亡机制的研究

院（部）系药学院

所学专业药学

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2013年 6 月 18 日

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20 年月日

摘要

目的观察蜂胶黄酮对过氧化氢（H2O2）诱导大鼠肾上腺嗜铬细胞瘤细胞（PC12）凋亡的影响及机制。

方法培养PC12细胞，取对数生长期细胞分为五组，空白对照组、模型组、蜂胶黄酮高、中、低剂量组，剂量分别为200mg/L、100 mg/L、50 mg/L.药物预处理2h后，孵育

H

2O

2

（140μmol/L）24h诱导过氧化损伤。TUNEL试剂盒检测原位细胞凋亡,流式细胞仪检测

细胞内活性氧水平以及细胞周期，ELISA检测细胞内Caspase-3蛋白含量。

结果TUNEL细胞凋亡染色显示, H2O2组与空白对照组比较染色明显加深,而蜂胶黄酮组染色明显变浅，说明蜂胶黄酮能对抗H2O2诱导细胞凋亡；周期结果显示H2O2组处于

G0/G1细胞明显增多，而处于G2/M、S期细胞明显减少，细胞增殖降低，而蜂胶黄酮增加

S期细胞促进细胞增殖；与空白对照组相比H

2O

2

组活性氧水平、细胞内Caspase-3含量明

显增高，而蜂胶黄酮各剂量组活性氧水平降低，Caspase-3含量减少。

结论蜂胶黄酮对H2O2诱发PC12神经细胞凋亡有显着的抑制作用，其机制可能其影响细胞周期、清除氧自由基、降低凋亡因子Caspase-3有关。

关键词蜂胶黄酮；PC12细胞；H2O2;Caspase-3；细胞凋亡

Abstract

Objective:

To observe the protection ofpropolis flavonoidson rat with injuried pheochromocytomacells(PC12) induced by?hydrogen peroxide(H2O2)

and to explore its possible mechanism.

Methods:Culture?PC12 cells,the cells inthe logarithmic growth phase weredivided into five groups,blank control group,model group,propolis flavoneof high,medium and low dose group,the dose of 200mg/L,100 mg/L,50 mg/L.After 2h 0fPharmacological preconditioning,the cellswere incubated with H2O2(140 μ mol/L)for24hto induceoxidative damage.TUNEL Kitis used fordetermining Cell apoptosis,Flow cytometry was used to detect the intracellular ROS level and cell cycle,ELISA is used for determining the concentration of protein Caspase-3 in cells. Results:Apoptosis of TUNEL cells staining, H2O2 group compared with the control group, staining was deepened, and the propolis flavone group was significantly lighter, that propolis flavonoids can antagonize the apoptosis induced by H2O2 Cycle showed that H2O2 group in

G0/G1 cells were increased, and in the G2/M,S phase cells decreased significantly, reduced cell proliferation, and propolis flavonoids increased S phase cells promoting cell proliferation; compared with the blank control group, caspase-3 in H2O2group, the levels of reactive oxygen species in cells increased significantly, while the propolis flavone in all dose groups decrease the levels of reactive oxygen species, reduced caspase-3 content.

Conclusion: Propolis flavonoids on H2O2-induced PC12 cells damage a significant protective effect,The mechanism may be its effect on cell cycle, scavenging oxygen free radicals, decrease the apoptosis related factor caspase-3.

Key words: Propolis; flavonoids; PC12 cells; H2O2; apoptosis